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30 November 2023, Volume 43 Issue 6

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INVITED REVIEWS

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Advances in reproductive isolation mechanisms during animal speciation

ZHANG Yuxing, WU Hong, YU Li

2023, 43(6):  627-643.  DOI: 10.16829/j.slxb.150838

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The process of speciation and its mechanism is not only the‘mystery of mysteries’of Darwin, but also a key to understanding the biological diversity in nature, and it is also one of the scientific focuses in the field of evolutionary biology. Speciation refers to the evolutionary process in which a new species diverges from its ancestors, and it is also the process of reproductive isolation or barrier establishment among populations. The exploration and revelation of this process and its inherent genetic mechanism are the key to understanding the phenomenon of biodiversity. By focusing on animal groups, this review systematically describes the ways of reproductive isolation establishment and their molecular mechanisms, then discusses the bottlenecks, challenges and potential opportunities faced by current research on animal reproductive isolations, with a view to providing reference and inspiration for subsequent research on animal speciation.

ORIGINAL PAPERS

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Population genomic analysis revealed the genetic basis of adaptations to urbanization in the lesser short-nosed fruit bat (Cynopterus brachyotis)

WANG Fei, ZHAO Huabin

2023, 43(6):  644-651.  DOI: 10.16829/j.slxb.150824

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The lesser short-nosed fruit bat (Cynopterus brachyotis) is a bat species commonly found in urban areas. A published study showed that rapid urbanization has led to a significant decrease in genetic diversity in this fruit bat. However, the genetic mechanisms underlying this species’ adaptations to urban environments have not been investigated. Based on the published population genome resequencing data from historical samples collected in 1931 and contemporary samples from 2011 - 2012, this study investigated the effects of rapid urbanization on the lesser shortnosed fruit bat and the genetic basis of adaptations to urbanization, using four methods (allele frequency analysis, selection signal detection based on population genetic differentiation, selection signal detection based on haplotypes, and detection of selected loci). Our results show that 669 selected genes were identified in the historical population and these genes were functionally enriched in visual and olfactory pathways. In contrast, 2 013 selected genes were identified in the contemporary population and these genes were enriched in immune, gustatory, and auditory pathways in addition to visual and olfactory-related pathways. These results suggest that rapid urbanization may have affected multiple sensory systems and immune systems in this species. Therefore, adaptive molecular evolution of immune, gustatory, and auditory genes may have occurred in C. brachyotis in response to rapid urbanization.

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The current distribution and prediction of suitable habitat of Eurasian lynx (Lynx lynx) in China

LIU Ke, LIU Yanlin, LI Sheng

2023, 43(6):  652-663.  DOI: 10.16829/j.slxb.150801

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Eurasian lynx (Lynx lynx) is widely distributed in northern Eurasia and, is one of the top predators in arid and semi-arid ecosystems in China and plays a significant role in maintaining ecosystem balance. Since the mid-20th century, the Eurasian lynx in China has experienced dramatic population decline and range contraction. However, our knowledge on its distribution status and dynamics is still very limited, which is a critical obstacle to developing specific conservation planning and action. To determine the current distribution of Eurasian lynx in China, we conducted a comprehensive search from multiple sources, including camera-trapping datasets, literature databases, and news reports, on the occurrence records of this species in China from 2010 to 2022. A total of 155 confirmed occurrence records of Eurasian lynx were obtained and were used to construct a species distribution model to predict its suitable habitat across China using MaxEnt with 24 environmental variables of 4 categories. The results showed that Eurasian lynx is currently distributed in 9 provinces, among which Neimenggu has the most occurrence records (n = 28). The result of cluster analysis showed that these occurrence sites could be grouped into three distinct spatial clusters: northeast, northwest, and southwest China. The averaged model AICc was the lowest when the regularization multiplier (RM) was 4 for the MaxEnt model and the mean AUC of the 10 candidate models was 0.814, indicating good performance of model prediction. The total area of the model predicted suitable habitat in China is 328.74×10⁴ km², mainly located in northern Heilongjiang, northeastern Neimenggu, northwestern Sichuan, eastern Xizang, Altai, Tianshan and Kunlun Mountains in Xinjiang, and the Qilian Mountains along the border of Qinghai and Gansu. Annual mean temperature, precipitation of driest month and land cover type were the most important factors affecting the habitat suitability of Eurasian lynx in China. Compared to the species range map provided by IUCN, the Tianshan-Altai Mountain Region, northeastern Neimenggu, parts of western Sichuan, and the Qilian Mountains were probably omitted during the IUCN species assessment, and further studies are urgently needed to examine the population and distribution status of Eurasian lynx in these regions.

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Distribution of suitable habitat for ungulates in Fanjingshan National Nature Reserve, Guizhou Province

MENG Bingshun, HUANG Xiaolong, XIE Bo, WANG Weixi, HUANG Jichi, ZHANG Tao, RAN Jingcheng, ZHANG Mingming

2023, 43(6):  664-675.  DOI: 10.16829/j.slxb.150756

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The spatial distribution of species and habitat suitability play an important role in the research on interspecific relationships, as well as research on relationships between species and environment. We evaluated and predicted the suitable habitats of four ungulates, Elaphodus cephalophus, Muntiacus reevesi, Pricornis milneedwardsii and Sus scrofa, distributed in Fanjingshan National Nature Reserve. Species occurrence data was based on the monitoring data by infrared cameras deployed in the study area from 2017 to 2022 and we used MaxEnt to model the suitable habitat for each species. The results showed that (1) the suitable habitats for Elaphodus cephalophus were mainly distributed in the eastern part of the reserve, covering an area of 19 993.17 hm², accounting for 46.1% of the protected area. The suitable habitats for Muntiacus reevesi covered an area of 15 470.97 hm² ( 35.6% of the protected area), distributed in the northern and southern parts of the reserve. The suitable habitats for Capricornis milneedwardsii were mainly distributed in the northern and eastern parts of the reserve, covering an area of 13 884.78 hm² (32.0% of the protected area), and the suitable habitats for Sus scrofa covered an area of 20 339.98 hm² (46.9% of the protected area), mainly distributed in the northern, eastern and southern parts of the reserve. (2) The suitable habitats of all four ungulates were mainly distributed in the core area of Fanjingshan Reserve. (3) The distribution of Elaphodus cephalophus was mainly influenced by altitude and vegetation type and the distribution of Capricornis milneedwardsii was mainly influenced by altitude, mean diurnal air temperature range and vegetation type. The characteristics of suitable habitats for Muntiacus reevesi and Sus scrofa were relatively similar, affected by mean diurnal air temperature range, isothermality and slope. In general, the distribution of suitable habitats for these four ungulates in Fanjingshan Reserve was the result of the interaction among the biological properties of species, natural environment, human activities and interspecific relationships. These results provide some insight into habitat suitability characteristics and influencing factors of ungulates in Fanjingshan National Nature Reserve.

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Seasonal differences in habitat selection of rhesus macaques (Macaca mulatta) in the Western Sichuan Plateau region

SI Huangkai, JIN Zuxiang, ZHANG Kechu, ZHOU Fei, YAO Yongfang, XIAO Hongtao, LI Bajin, Pucuowangjia, XU Huailiang

2023, 43(6):  676-688.  DOI: 10.16829/j.slxb.150752

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From July 2019 to March 2021, a follow-up survey was conducted on a group of about 40 rhesus macaques (Macaca mulatta) in Yajiang County, Sichuan Province, to explore seasonal differences in habitat selection in a high-altitude environment. 135 plots (20 m×20 m) were selected based on macaque observations and 60 control plots were set where macaques were not present. 18 ecological factors, such as altitude, grazing intensity, and vegetation types, were measured in each plot and used to calculate a selection index and perform a principal component analysis. The results showed that (1) rhesus macaques preferred staying at higher altitudes (3 900 - 4 300 m) in spring, summer and autumn, but moved toward lower altitudes (3 500 - 3 900 m) in winter; (2) the macaques preferred sunny slopes, while in winter, there was no obvious preference for slope orientation; (3) their preferred habitats were areas with high canopy density (greater than 60%) in spring and low canopy density (less than 60%) in autumn and winter; (4) in spring and winter, they stayed near water sources (less than 100 m away), but preferred habitats were more than 100 m from water sources in summer and autumn; (5) in summer, they preferred to move to forested areas (coniferous forest, alpine oak forest, mixed broadleaf-conifer forest), but in other seasons they favored lowland grasslands (near the river valley) at lower altitude; (6) they avoided areas with high levels of human interference as much as possible in summer and autumn and engaged in more activities in areas with high levels of human interference in winter and spring. Both human disturbance and geographical factors played a significant role in year-round habitat selection. Vegetation factors mainly influenced habitat selection in spring, summer, and autumn, while shelter condition factors mainly affected summer habitat selection. Compared to other low-altitude areas, maximizing access to food and reducing energy consumption are the key factors for macaques to survive and reproduce in plateau environments with low natural productivity.

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Feeding behavior of sympatric rodents in the Alxa Desert in response to different grazing patterns

LIU Lina, YUAN Shuai, LI Xin, FU Heping, WU Xiaodong

2023, 43(6):  689-701.  DOI: 10.16829/j.slxb.150800

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The fragile ecosystem of the Alxa Desert is sensitive to human disturbance and different grazing patterns lead to different vegetation resource types, density, distribution, coverage, substrate and animal perception of predation risk. The feeding differences of desert sympatric rodents under different grazing disturbances are of great significance for the maintenance of biodiversity in desert areas. In 2020, infrared camera trap technology was used to observe the feeding behavior of three dominant rodent species, namely Dipus sagitta, Orientallactaga sibirica, and Meriones meridianus, which coexisted in the same area in the Alax Desert. The results showed that: (1) the population density of the dominant rodent species was positively related to their feeding behavior under different grazing patterns. Grazing increased O. sibirica population density, and its feeding behavior also accounted for more in grazing-intensive habitats. M. meridianus had higher population density and more feeding behavior in habitats with low grazing intensity and high vegetation coverage. D. sagitta devoted more foraging effort in more grazing-intensive habitats. (2) Under the same grazing mode, M. meridianus adopted a combined feeding strategy of ‘foraging + feeding + storage’ , while D. sagitta and O. sibirica adopted the feeding strategy of ‘foraging + feeding’ . These results indicated that the feeding behaviors of rodent species with various feeding strategies were affected by grazing patterns according to changing environmental factors such as rodent population density, habitat vegetation, and soil.

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Preliminary analysis of the molecular phylogeography of Sorex unguiculatus and Sorex gracillimus

MEI Xiufeng, CAI He, HAN Meifeng, ZHAO Xinxu, ZHANG Chunfeng, SHI Yanjing, CHEN Huan, TIAN Xinmin, ZHANG Junsheng, LIU Zhu

2023, 43(6):  702-709.  DOI: 10.16829/j.slxb.150699

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In order to clarify the population phylogeographic relationship and subspecies differentiation of Sorex unguiculatus and Sorex gracillimus, this study analyzed the Cyt b gene sequences of S. unguiculatus (11 samples) and S. gracillimus (17 samples) collected from Northeast China together with Cyt b gene sequences of S. unguiculatus (32 samples) and S. gracillimus (22 samples) downloaded from GenBank. The different geographical populations of the two species showed different geographical patterns. In the phylogenetic tree of S. unguiculatus, there was no correlation between molecular system relation and geographical distribution. In the phylogenetic tree of S. gracillimus, samples from Northeast China were clustered into a separate clade apart from samples from Russia and Hokkaido, Japan. A similar geographical pattern was also observed in the Median-joining network analysis. Neutral tests supported S. unguiculatus has experienced population expansions in the recent past, but S. gracillimus has not experienced dramatic population expansions. The results also supported that S. unguiculatus has no subspecies differentiation. On the other hand, we found that S. gracillimus was divided into two subspecies across the geographic area of this study, one subspecies distributed in Northeast China and the other subspecies distributed in the Russian Far East (from southern Magadan to Primorsky Krai, Sakhalin Island included) and Hokkaido, Japan.

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Effect of exposure to high temperature on energy metabolism and oxidative stress in striped hamsters

ZHANG Ruihan, LUO Dan, LUO Xinxin, HU Chenxiao, LIN Long, CHEN Qian, CAO Jing, ZHAO Zhijun

2023, 43(6):  710-722.  DOI: 10.16829/j.slxb.150822

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The adaptive regulation of animal energy metabolism affects life history characteristics such as growth, development, reproduction, and aging. Metabolic rate and tissue mitochondrial respiration rate are related to free radical levels, which are important factors influencing the aging of the organism. In this study, we investigated the intrinsic link among energy metabolism, tissue respiration rates of major metabolically active organs, free radical levels, and antioxidant capacity in response to high ambient temperature. We used as a model species striped hamsters (Cricetulus barabensis) that were previously acclimated to room temperature (21 ℃) and warm temperature (32.5 ℃) for 4 weeks and then were acutely exposed to extremely high temperature (37 ℃) for 48 h. The energy intake, metabolic rate, body temperature, mitochondrial respiration rate, and uncoupling protein (UCP) gene (ucp) expression of brown adipose tissue (BAT), liver and skeletal muscle were measured. The levels of protein carbonyl and malondialdehyde (MDA), and the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were also determined. The results showed that the acclimation to 32.5℃ and acute exposure to 37℃ resulted in significant reductions in gross and digestive energy intake, basal metabolic rate, and non-shivering thermogenesis. Cytochrome c oxidase (COX) activity was reduced after acclimation to 32.5℃ by 84.2%, 50.0%, and 56.6% in BAT, liver, and muscle, respectively. The liver COX activity decreased further by 23.3% after exposure to 37℃, but COX activity in BAT and muscle was not significantly changed. The acute exposure to 37℃ deceased BAT ucp1 expression by 89.5%, liver ucp2 by 76.2%, and muscle ucp3 by 58.8% in the hamster previously acclimated to 21℃, but it had no significant effect on the gene expression in the hamsters previously acclimated to 32.5℃. Acclimation to 32.5℃ significantly decreased MDA levels in heart, lung, kidney, and muscle, but not in liver and brain, and it significantly increased protein carbonyl levels in the kidney only. Acute exposure to 37℃ significantly reduced protein carbonyl levels in liver and heart, but it had no significant effect on MDA levels in all tissues. In addition, acclimation to 32.5℃ significantly reduced SOD activity in liver and muscle, but did not affect heart, lung, kidney, or brain; and it decreased GSH-Px activity in heart, lung, and kidney, but increased GSH-Px activity in muscle, and had no effect in liver. These findings suggest that (1) the effects of acute exposure to high temperatures on energy metabolism and mitochondrial respiration rate are related to habitat ambient temperature, with the animals at lower ambient temperature being more affected by high temperature; (2) after being acclimated to the warm and acutely exposed to high temperature, the metabolic rate of the organism and mitochondria respiratory decreased, while the liver, heart, lung, kidneys, skeletal muscle, and brain do not show considerable oxidative stress and damage, being inconsistent with the‘free radical hypothesis’ .

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The expression profile of miRNA in the different tissues of forest musk deer (Moschus berezovskii)

YU Miaojie, XU Zhongxian, JIANG Xuemei, WANG Chunhua, ZHAO Chanjuan, QI Wenhua, JIE Hang

2023, 43(6):  723-733.  DOI: 10.16829/j.slxb.150744

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Forest musk deer (Moschus berezovskii) is a medicinal economic species and the musk secreted by the male musk deer is an important component of Chinese patent medicine and perfume, which has high medicinal and economic value. miRNAs play an important role in animal cell development, differentiation, apoptosis, and disease development, which are selectively expressed in different tissues. The heart, liver, spleen, lung, kidney, and muscle tissues of forest musk deer were high-throughput sequenced by using RNA-seq technology on the Hi-seq 2500 sequencing platform. The microRNA expression of these 6 tissues was analyzed by using biostatistics and bioinformatics methods and RT-qPCR was used to verify the sequencing results. The results showed that 1 650 miRNAs co-expressed in the heart, liver, spleen, lung, kidney, and muscle tissues of forest musk deer, of which there were 86, 54, 44, 68, 83, and 50 specific miRNAs, respectively (P < 0.01). The expression levels of co-expressed miRNAs were significantly different in these six types of tissues (P < 0.05) and the expression levels of some specific miRNAs in the heart were much higher than that in other tissues. KEGG analysis showed that the target genes of specific miRNAs in the liver of forest musk deer were closely related to the Wnt signaling pathway (ko04310), cAMP signaling pathway (ko04024), ECM-receptor interaction (ko04512), Notch signaling pathway (ko04330), and AMPK signaling pathway (ko04152) (P < 0.01). Nine miRNAs were randomly selected for verification with RT-qPCR in liver tissue of health and abscess forest musk deer and the results were consistent with the sequencing results.

REVIEWS

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Application of deep learning in wildlife conservation

ZHONG Junjie, NIU Bing, CHEN Qin, CHEN Xiang, WANG Yan

2023, 43(6):  734-744.  DOI: 10.16829/j.slxb.150774

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Wildlife is one of the vital biological resources, but the increasing human activities and environmental degradation pose a severe threat to the survival of wild animals. Deep learning has emerged as a prominent research direction in the field of artificial intelligence and has been widely applied across various disciplines. Its versatility has demonstrated enormous potential in wildlife conservation, particularly in image recognition, monitoring, and audio recognition. This article introduces several common deep learning algorithms, provides an overview of the applications of different deep learning models in wildlife conservation, and analyzes the current issues and challenges, including limited training data, variability of environmental conditions, and the complexity of wildlife behavior. In the future, to employ deep learning for wildlife protection, in addition to addressing challenges such as data acquisition and utilization, and robustness in image recognition against various interferences, it is crucial to develop more robust and efficient deep learning models that cater to the specific requirements of wildlife conservation.

METHOD AND TECHNOLOGY

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Prokaryotic expression system construction, antibody preparation and tissues distribution of bactrian camel TLR1

XIE Dongxu, ZHANG Rui, Suonanji, LIU Kejiang, WANG Tingwei, WANG Wenhui

2023, 43(6):  745-752.  DOI: 10.16829/j.slxb.150780

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In this study, we used bioinformatics software to analyze the TLR1 (Toll-like receptors 1) gene and used a prokaryotic expression system to inducibly express TLR1 recombinant plasmids into TLR1 protein. Rabbit polyclonal antibactrian camel TLR1 antibody was generated using the expressed recombinant TLR1 protein, and the expression of TLR1 in organs including bactrian camel heart, liver, spleen, lung, and kidney was detected by HE staining coupled with immunohistochemical staining of SABC using the prepared TLR1 polyclonal antibody. The size of the protein of interest obtained in this study was 62 kDa, which was the same as predicted by bioinformatics software. TLR1 recombinant protein was mainly expressed in the form of inclusion bodies. In this study, the titer of the TLR1 antibody was determined by indirect ELISA, which showed that the antibody titer result was 1∶ 64 000, and the specificity of the TLR1 antibody was confirmed by Western blot analysis. The results of the present trial of HE staining with immunohistochemical staining of SABC showed that TLR1 was upregulated in cardiomyocytes of bactrian camel hearts, Kupffer cells of the liver, monocytes/macrophages of the spleen, the respiratory bronchiolar epithelium, and type Ⅰ and type Ⅱ alveolar cells of the lung. A positive reaction appeared on the epithelial cells of the distal convoluted tubule of the kidney. In conclusion, the rabbit anti-bactrian camel TLR1 polyclonal antibody has good specificity and can be used for immunohistochemical detection of TLR1 and provides a basis for further investigation of the function of TLR1 in different tissues and organs of bactrian camels.

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Dynamic observation of provisioned wild Trachypithecus francoisi in Mayanghe National Nature Reserve, Guizhou

ZHANG Peng, YAN Xiugang, SHI Yun, YANG De, WANG Bin, LI Wei, YANG Yuanlu, YANG Zhaohui, WU Guangyang, CAI Zhi, LI Siyao, ZOU Hao, ZHANG Fan

2023, 43(6):  753-759.  DOI: 10.16829/j.slxb.150707

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From 2009 to 2021, a direct observation method was used to track and monitor a provisioned group of Trachypithecus francoisi in Mayanghe National Nature Reserve. The observation period covered 1 846 days with an average annual working days of 142. In total, 31 adults (15 males, 16 females) were observed and, in addition, 26 babies were born with a male-female ratio of 1.2∶1 (14 male, 12 female) of which 15 infants survived (7 males, 8 females), a survival rate of 57.7%. For the whole group, the average annual monkey group birth rate was about 22.8% and the annual loss rate was about 11.4%. The number of births exhibited a bimodal distribution, peaking from February to April and from July to October. The average reproductive interval of adult monkeys was 18.7 months but shortened to 16.3 months after the death or disappearance of a newborn. During 2019 -2021, the average growth rate of the group was 11.4%, the number of monkeys had generally stabilized and was self-sustaining. However, as of the end of December 2021, the feeding group had dwindled to just nine individuals (two males and seven females) due to individual deaths, disappearances, and expulsions of some sub-adults. There were no infants or juveniles and insufficient sub-adults. Furthermore, the proportion of middle-aged and elderly individuals was high, resulting in an imbalanced age structure. This study provides basic data related to long-term monitoring of wild Trachypithecus francoisi in Mayanghe National Nature Reserve. This has provided an important basis for follow-up scientific research, educational projects, and monitoring and management of Trachypithecus francoisi in the reserve. Moreover, it has ensured that this flagship species and its habitats are effectively protected and sustainably used, so as to promote the high-quality development of biodiversity protection in this region.

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New records of Paguma larvata in Liaoning Province, China

ZHANG Lei, CHANG Jin, FENG Chaofan, WAN Dongmei

2023, 43(6):  760-700.  DOI: 10.16829/j.slxb.150815

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Paguma larvata is mainly distributed in the central and southern regions in China. It is listed as a near-threatened species in the Red List of China’ s Vertebrates. In the past, there were no records of distribution of this species Liaoning Province. During the camera trapping survey of animal diversity in the Qinglonghe, Bailangshan, and Wuhuading National Nature Reserves in the western region of Liaoning, 215 independent detections of Paguma larvata were captured by camera-trapping with 584 photographs and 106 video clips from January 2022 to May 2023. These image data represent the first record of Paguma larvata in Liaoning Province, which not only plays an important role in the study of biodiversity and species integrity in this province but also provides the basic data for the distribution of Paguma larvata in China.